Figure 2
From: Deoxyribonuclease 1 reduces pathogenic effects of cigarette smoke exposure in the lung
Cigarette smoke exposure induces formation of neutrophil extracellular traps. CSE induces neutrophil extracellular trap (NET) formation in human peripheral blood neutrophils in vitro (A–D). (A) Representative image shows NET formation as characterised by staining for extracellular chromatin, H3Cit, peptidylarginase deiminase (PAD) 4 and neutrophil elastase (NE). Scale bar = 20μm. (B) NET expression in blood neutrophils (n = 8 individuals). (C) A flow cytometry method was also developed to measure NET production, with SYTOX green staining for extracellular chromatin production and eFluor as a marker of cell death. Dot plot shows NET expression as characterised by positive staining for SYTOX green and negative for eFlour. (D) NET expression by neutrophils demonstrated using flow cytometry (n = 8). Exposure to cigarette smoke induces NET expression in mice in vivo (E,F). (E) Image shows NET formation in lung tissue. Scale bar = 10μm. (F) NET expression in lung tissue (n = 10). (G) In lung tissue, cigarette smoke induced a higher number of METs than NETs.
